EP3234194A1 - New method for detecting human butyrylcholinesterase - Google Patents
New method for detecting human butyrylcholinesteraseInfo
- Publication number
- EP3234194A1 EP3234194A1 EP15834679.1A EP15834679A EP3234194A1 EP 3234194 A1 EP3234194 A1 EP 3234194A1 EP 15834679 A EP15834679 A EP 15834679A EP 3234194 A1 EP3234194 A1 EP 3234194A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- optical resonator
- biosensor
- resonator biosensor
- antibody
- probe
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims description 16
- 101000943274 Homo sapiens Cholinesterase Proteins 0.000 title claims description 9
- 102000051276 human BCHE Human genes 0.000 title claims description 9
- 239000000523 sample Substances 0.000 claims abstract description 70
- 230000003287 optical effect Effects 0.000 claims abstract description 67
- 102000021944 Butyrylcholinesterase Human genes 0.000 claims abstract description 20
- 108010053652 Butyrylcholinesterase Proteins 0.000 claims abstract description 20
- 238000001514 detection method Methods 0.000 claims abstract description 16
- 150000002903 organophosphorus compounds Chemical class 0.000 claims description 12
- 150000001875 compounds Chemical class 0.000 claims description 10
- 125000004432 carbon atom Chemical group C* 0.000 claims description 9
- 125000006850 spacer group Chemical group 0.000 claims description 7
- 125000003545 alkoxy group Chemical group 0.000 claims description 3
- 125000004093 cyano group Chemical group *C#N 0.000 claims description 3
- 229910052736 halogen Inorganic materials 0.000 claims description 3
- 150000002367 halogens Chemical class 0.000 claims description 3
- 125000005842 heteroatom Chemical group 0.000 claims description 3
- 125000001183 hydrocarbyl group Chemical group 0.000 claims description 3
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- 229910052760 oxygen Inorganic materials 0.000 claims description 3
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 3
- 238000006467 substitution reaction Methods 0.000 claims description 3
- 229910052717 sulfur Inorganic materials 0.000 claims description 3
- 125000004169 (C1-C6) alkyl group Chemical group 0.000 claims description 2
- 230000027455 binding Effects 0.000 description 14
- 229940088598 enzyme Drugs 0.000 description 11
- 102000004190 Enzymes Human genes 0.000 description 10
- 108090000790 Enzymes Proteins 0.000 description 10
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical group N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 9
- 230000005540 biological transmission Effects 0.000 description 8
- 239000012491 analyte Substances 0.000 description 5
- 229940058344 antitrematodals organophosphorous compound Drugs 0.000 description 5
- 238000003556 assay Methods 0.000 description 5
- 229960002685 biotin Drugs 0.000 description 5
- 235000020958 biotin Nutrition 0.000 description 5
- 239000011616 biotin Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 239000003958 nerve gas Substances 0.000 description 4
- 239000000575 pesticide Substances 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 108010090804 Streptavidin Proteins 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 238000002032 lab-on-a-chip Methods 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 238000012544 monitoring process Methods 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 102100033639 Acetylcholinesterase Human genes 0.000 description 2
- 108010022752 Acetylcholinesterase Proteins 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000004590 computer program Methods 0.000 description 2
- 231100000673 dose–response relationship Toxicity 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
- 108090001008 Avidin Proteins 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 102000003914 Cholinesterases Human genes 0.000 description 1
- 108090000322 Cholinesterases Proteins 0.000 description 1
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical group OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 1
- 229920005439 Perspex® Polymers 0.000 description 1
- DYAHQFWOVKZOOW-UHFFFAOYSA-N Sarin Chemical compound CC(C)OP(C)(F)=O DYAHQFWOVKZOOW-UHFFFAOYSA-N 0.000 description 1
- 238000003848 UV Light-Curing Methods 0.000 description 1
- JJIUCEJQJXNMHV-UHFFFAOYSA-N VX nerve agent Chemical compound CCOP(C)(=O)SCCN(C(C)C)C(C)C JJIUCEJQJXNMHV-UHFFFAOYSA-N 0.000 description 1
- 206010052428 Wound Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- PJVJTCIRVMBVIA-JTQLQIEISA-N [dimethylamino(ethoxy)phosphoryl]formonitrile Chemical compound CCO[P@@](=O)(C#N)N(C)C PJVJTCIRVMBVIA-JTQLQIEISA-N 0.000 description 1
- 229940022698 acetylcholinesterase Drugs 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229940048961 cholinesterase Drugs 0.000 description 1
- 239000000544 cholinesterase inhibitor Substances 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000000779 depleting effect Effects 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000007824 enzymatic assay Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000002917 insecticide Substances 0.000 description 1
- 231100000566 intoxication Toxicity 0.000 description 1
- 230000035987 intoxication Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 description 1
- 125000006216 methylsulfinyl group Chemical group [H]C([H])([H])S(*)=O 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000009149 molecular binding Effects 0.000 description 1
- 239000013307 optical fiber Substances 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 239000004038 photonic crystal Substances 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 239000004926 polymethyl methacrylate Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012207 quantitative assay Methods 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 108700012359 toxins Proteins 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000010626 work up procedure Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/916—Hydrolases (3) acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)
- G01N2333/918—Carboxylic ester hydrolases (3.1.1)
Definitions
- the invention relates to the field of detection of biological compounds, particularly for the detection of butyrylcholinesterase. Such a detection may be advantageously used in the detection of the duration and severity of exposure to cholinesterase inhibiting nerve gases or pesticides.
- Organophosphorous compounds are a group of the most toxic compounds that are known. Many of those have been used in warfare as nerve gas, such as sarin, tabun and VX. Organophosphorous toxins are also used as pesticides and insecticides.
- optical resonator biosensor for the detection of butyrylcholinesterase comprising two optical resonator biosensor systems in which in the first optical resonator biosensor system a probe is attached to the resonator wherein said probe is able to bind butyrylcholinesterase, preferably wherein said probe is able to bind uninhibited butyrylcholinesterase.
- the optical resonator biosensor provides in the second optical resonator biosensor system an antibody that is attached to the resonator, wherein said antibody is able to bind butyrylcholinesterase, preferably wherein said antibody is able to bind both inhibited and uninhibited
- butyrylcholinester ase butyrylcholinester ase .
- the optical resonator biosensor is provide with a sample, applied in such a way that it comes into contact with both optical resonator biosensor systems., preferably by applyingthe the sample via a capillary track.
- both optical resonator biosensor systems are placed along one capillary. In such a system preferably the amount of probes in said first optical resonator biosensor is sufficient large to capture all the uninhibited butyrylcholinesterase.
- biosensor wherein one optical resonator biosensor system is placed along one branch of the capillary, while the other optical resonator biosensor system is placed along an other branch of the capillary.
- a further preferred embodiment is a biosensor wherein the resonator is a ring resonator.
- Another further preferred embodiment is a biosensor wherein the resonator biosensor is a waveguide, preferably wherein said waveguide is part of an interferometer.
- a further part of the invention is a method for determining the exposure to an organophosphorous compound by measuring the amounts of inhibited, uninhibited and total butyrylcholinesterase in a sample by assaying said sample with an optical resonator biosensor according to the invention.
- optical resonator biosensor for the determination of the exposure to an organophosphorous compound, wherein said optical resonator biosensor is an optical resonator biosensor according to the invention.
- optical resonator biosensor method and use the butyrylcholinesterase is a human butyrylcholinesterase. Further, in the above mentioned optical resonator biosensor, method and use the probe o
- R is an spacer chain with more than 10 C-atoms, which is able to be bound to the sensor surface.
- said spacer chain is a substituted or unsubstituted, linear or branched hydrocarbon chain of more than 10 carbon atoms, wherein one to three of the C atoms may be replaced by a heteroatom selected from the group of O, N and S, and wherein the chain may have substitutions selected from the group of halogen, phenyl, linear or branched O-C1-C6 alkyl, linear or branched alkoxy, nitro, cyano or methylsulfinyl. Examples of such probes are:
- the antibody that binds to butyrylcholinesterase in the above mentioned optical resonator biosensor, method and use is monoclonal antibody 3E8.
- Fig. 1 shows the dose-dependent shift of resonsnace wavelength from various doses of human butyrylcholinesterase applied to a biosensor on a ring resonator.
- the Y-axis denots the shift in wavelength, while the x-axis shows the time in seconds.
- Butyrylcholinesterase is an enzyme that is inhibited when an organosphosphorous compound, such as a nerve gas or a pesticide, binds to its active site. Inhibition of BuChE in itself does not give any toxic effects (in comparison to inhibition of acetylcholinesterase, AChE, which causes the toxicity demonstrated by the organophosphorous compounds), but it can ideally be used as a biomarker.
- the concentration of inhibited BuChE in comparison with the total BuChE concentration conveys information whether or not a subject has been exposed to an organophosphorous compound, as well as an indication of the extent of the exposure.
- an antibody that is able to bind to the enzyme is used.
- the amount of inhibited BuChE is the total amount of BuChE minus the amount of uninhibited BuChE.
- the word "probe” is used for a molecule that is able to bind uninhibited BuChE.
- an uninhibited BuChE' of the present invention is a molecule wherein the recognition site for an
- organophosphorous compound has not been occupied, which in general means that the enzyme is still able to exert its enzymatic action. Further, the 'inhibited' or 'uninhibited' state is not altered by binding to the ant- BuChE antibody as defined herein.
- an anti-BuChE antibody is preferably a monoclonal antibody that is able to bind to both inhibited and uninhibited BuChE.
- Such antibodies are known in the art (e.g. Wang, L. et al., 2011, Analytica
- an optical detection method an optical resonator biosensor or ring resonator.
- An optical resonator biosensor works through monitoring resonance wavelength shifts which occur in optical response of the ring resonator, i.e. its transmission as function of
- An optical resonator biosensor consists of three parts: 1) a biorecognition element, which in the present invention is a probe or an antibody that is able to bind BuChE, more preferably human BuChE; 2) an optical transducer (the optical resonator) and 3) an electronic readout scheme for measuring and recording optical frequency shifts (Vollmer, F. and Baaske, M. 2012, ChemPhysChem 13:427- 436).
- the optical resonator may be in the form of a glass microsphere, but may also be formed by micro-capillaries and liquid-core optical ring resonators (LCORR), micro-toroids, micro-disks, photonic crystal cavities and micro-rings.
- LCDORR liquid-core optical ring resonators
- a ring resonator is used.
- the general lay-out of such micro-rings is known in the art (e.g. Lin, S.Y. et al., 2010, Nano Lett. 10:2408-2411; Nitkowski A. et al., 2011, Biomed. Optics Express 2:271-277; Carlborg, C.F. et al., 2010, Lab on a Chip 10:281-290).
- optical resonator biosensor a complete test system can be obtained on a very small scale: the actual measuring device can be provided as a lab-on-a-chip technology, where e.g. a ring resonator maybe used with a diameter of 5- 100 pm. This allows testing in the field.
- the speed of detection can be very high (within a few seconds to a few minutes), while the amount of material needed, the sample volume, may be minimal (in the nanoliter or microliter range).
- the optical resonator biosensor systems usually combine a very high sensitivity with a good specificity.
- a sample may be obtained from any body fluid that contains the enzyme butyrylcholinesterase, but preferably a blood sample. Since only a small amount of sample is needed to be applied to the testing device, blood from a finger prick is an ideal source, since it is readily obtained even under field conditions. However, also blood samples from other sources (e.g.
- venipuncture, or bleeding wounds may be used.
- the optical resonator biosensor in the present invention at least needs to measure the uninhibited BuChE level.
- the resonator is provided with a probe that is able to bind uninhibited BuChe, more preferably uninhibited human BuChE (HuBuChE).
- This probe advantageously is a compound that is able to bind to the BuChe at the active site, i.e. a compound that resembles an organophosphorous compound as defined above or any other molecule that is able to bind to the active site of the enzyme.
- this is a compound with the general formula:
- R is an spacer chain with more than 10 C-atoms, which is able to be bound to the sensor surface.
- said spacer chain is a substituted or unsubstituted, linear or branched hydrocarbon chain of more than 10 carbon atoms, wherein one to three of the C atoms may be replaced by a heteroatom selected from the group of O, N and S, and wherein the chain may have substitutions selected from the group of halogen, phenyl, linear or branched ⁇ -Ci-Ce alkyl, linear or branched alkoxy, nitro, cyano or
- the chain may comprise a (poly)ethylene glycol chain.
- the part of R that is capable of binding to the sensor is a biotin moiety or an amino moiety.
- the sensor In the fall of a biotin moiety, the sensor binds with an avidin moiety at the surface of the sensor.
- an amino moiety In the fall of an amino moiety, said moiety binds with a reactive carboxyl-group that is present at the surface of the biosensor.
- the skilled person will know how to provide the biosensor surface with the above-mentioned components for binding with their counterparts on the probe.
- the R group may vary considerably and primarily acts as a spacer to prevent interaction of the bound enzyme with the biosensor.
- an aqueous composition comprising free probe is added to the sample on the sensor.
- the probe may first bind with the enzyme and then together with the enzyme may be bound to the sensor surface. Whether or not the actual binding to the enzyme in such a situation takes place before or after the probe has attached itself to the sensor surface is not relevant.
- a refractive index sensor such as a ring resonator
- HuBuChE or the binding of the enzyme-antibody or enzyme-probe complex can be detected as refractive index variations by monitoring the shift of the resonance wavelength as a function of time. Since the amount of shift corresponds to the amount of molecules bound, this provides a reliable, quick and a quantitative detection of the amount of uninhibited BuChE.
- BuChE BuChE
- the antibody may be bound to the surface of the sensor or may first react freely in the test (sample) solution with the BuChE and after reaction as antibody-enzyme complex be bound to the sensor.
- the skilled person is capable of finding ways how to bind the antibody to the sensor (e.g. by immobilising on the sensor an antibody that recognizes the anti-BuChe-antibody).
- An example of an antibody that is able to bind to both inhibited BuChE and uninhibited BuChE may be the monoclonal antibody 3E8, which is commercially available (e.g. from
- a sample is provided to a sample inlet after which the sample flows in two capillaries each to a different sensor.
- the signals obtained from those sensors are processed to calculate the
- the two sensors are placed along one and the same capillary that runs from the sample inlet site.
- the sample flows across one sensor to measure the concentration of one analyte, and the same sample subsequently flows across the second sensor to measure the other analyte.
- the probe and/or the antibody may be available as free soluble in the sample solution or bound to the sensor surface.
- the signals from both sensors are then further processed to ultimately yield the concentration and/or percentage of inhibited BuChE in the sample.
- Such a sequential arrangement is allowed when the first sensor only binds a small fraction of the analyte (not depeleting the sample) such that the total concentration of analyte in the sample is not markedly affected.
- the total amount of BuChE is measured by one sensor in one branch of the capillary, while in a second branch of the capillary all uninhibited BuChE is captured by a gel in which an excess amount of probe is available. After capturing all the uninhibited BuChE from the sample, the remaining amount of BuChE, which then necessarily is inhibited BuChE, is measured with immobilized antibodies.
- the first measurement in the first capillary may also be a measurement of the amount of uninhibited BuChE, since the total amount of BuChE, and thus the percentage of inhibited BuChE, can be calculated in either way.
- the amount of uninhibited BuChE is measured quantitatively, while still depleting the sample from this uninhibited BuChE. This can be accomplished by having an excess amount of probe on the sensor. Since, beforehand it is not known how high the concentration of uninhibited BuChE will be, it would be possible to accommodate a dilution series of the sample to be measured by a series of sensors, where the sample is split and diluted in such a way that there is at least one sensor which would contain an excess amount of probe in order to completely delete this (diluted) sample.
- the analysis device may be a lab-on-a-chip, which has a sample inlet site at which a sample to be assayed, such as a drop of blood, is applied.
- a further aqueous solution containing a free soluble probe or antibody as described above may be added.
- this 'chip' has one, two or more capillaries in which the sample is further guided to the one, two or more sensors at which the analyte is eventually detected.
- the signals that are obtained from these, one, two or more sensors are being sent to a calculating unit, which calculating unit may be a micro-computer, which is situated on the chip itself, or the calculating unit may be removed from the sensors and the results are transmitted.
- the signals can be stored in a storage medium on the chip and can be read out when contacting the chip with a transmission unit or directly with a calculating unit.
- Contacting the chip with a transmission device or calculating device may be in the form of any kind of contact that is able to transfer the signals, such as cables, a bus-like structure, such as a USB-stick or the like.
- the signals may be transmitted directly by a wireless transmitter, which may be any kind of wireless transmission system, i.e. modulated or unmodulated, encrypted or non-encrypted, radiosignals, optic, sonic or electromagnetic, etc.
- a wireless transmission system is a WiFi system or an infra-red system.
- the biosensor system also provides an indicator which generates a signal when an excess amount of inhibited BuChE has been detected.
- an indicator can provide a chemical, electrical, optical or acoustical signal.
- the signal may be directly produced by one or more of the reagents contained in the system, but it may also be produced indirectly through input from the calculating unit.
- aspects of the present invention may be embodied as a system, method or computer program product.
- aspects of the present invention may take the form of an entirely hardware embodiment, or an embodiment combining software and hardware aspects that may all generally be referred to herein as a "circuit,” “module” or “system.”
- aspects of the present invention may take the form of a computer program product embodied in one or more computer readable medium(s) having computer readable program code embodied thereon.
- Program code embodied on a computer readable medium may be transmitted using any appropriate medium, including, but not limited to, wireless, wireline, optical fiber cable, RF, etc., or any suitable combination of the foregoing.
- streptavidin 40 ug/ml in phosphate buffer saline
- a solution with biotinylated antibodies (3E8, 1 ug/ml in phosphate buffer saline) was applied such that the antibody was able to bind to the streptavidin.
- the integrated optical chip carries a ring resonator.
- the ring resonator has a length of 500 um.
- a tunable laser was continuously swept in steps of 1 pm across 4 nm and for each sweep the transmission of the ring was recorded as function of applied wavelength. From the recorded spectra over time, the shift of the resonance wavelengths as result of refractive index changes caused by human butyrylcholinesterase binding to the antibodies, is determined and plotted as function of time.
- This experiment was performed for different concentrations of human butyrylcholinesterase in phosphate buffer saline, that were applied to the sensor at a flow rate of 30 ul/min. As is shown in Fig. 1 a dose-dependent shift in resonance
Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP14198901.2A EP3034621A1 (en) | 2014-12-18 | 2014-12-18 | New method for detecting human butyrylcholinesterase |
PCT/NL2015/050883 WO2016099275A1 (en) | 2014-12-18 | 2015-12-18 | New method for detecting human butyrylcholinesterase |
Publications (2)
Publication Number | Publication Date |
---|---|
EP3234194A1 true EP3234194A1 (en) | 2017-10-25 |
EP3234194B1 EP3234194B1 (en) | 2020-09-09 |
Family
ID=52347092
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP14198901.2A Ceased EP3034621A1 (en) | 2014-12-18 | 2014-12-18 | New method for detecting human butyrylcholinesterase |
EP15834679.1A Active EP3234194B1 (en) | 2014-12-18 | 2015-12-18 | New method for detecting human butyrylcholinesterase |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP14198901.2A Ceased EP3034621A1 (en) | 2014-12-18 | 2014-12-18 | New method for detecting human butyrylcholinesterase |
Country Status (3)
Country | Link |
---|---|
US (1) | US20170356913A1 (en) |
EP (2) | EP3034621A1 (en) |
WO (1) | WO2016099275A1 (en) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108192597B (en) * | 2016-12-08 | 2019-07-12 | 华中师范大学 | Half cyanines class fluorescence probe of near-infrared and its preparation method and application for detecting butyrylcholine esterase |
CN110294720B (en) * | 2018-03-23 | 2021-05-07 | 香港理工大学深圳研究院 | Fluorescent probe for detecting butyrylcholine esterase and application thereof |
EP3663297A1 (en) | 2018-12-06 | 2020-06-10 | Nederlandse Organisatie voor toegepast- natuurwetenschappelijk onderzoek TNO | Diels-alder reaction with furanics to obtain aromatics |
EP3844165A1 (en) | 2018-08-29 | 2021-07-07 | Nederlandse Organisatie Voor Toegepast- Natuurwetenschappelijk Onderzoek Tno | Diels-alder reaction with furanics to obtain aromatics |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7796262B1 (en) * | 2007-05-31 | 2010-09-14 | Nomadics, Inc. | Integrated optical resonator device for measuring chemical and biological analyte concentrations |
US20110166043A1 (en) * | 2007-11-14 | 2011-07-07 | Jon Owen Nagy | Biomarker detection-2 |
-
2014
- 2014-12-18 EP EP14198901.2A patent/EP3034621A1/en not_active Ceased
-
2015
- 2015-12-18 WO PCT/NL2015/050883 patent/WO2016099275A1/en active Application Filing
- 2015-12-18 US US15/537,636 patent/US20170356913A1/en not_active Abandoned
- 2015-12-18 EP EP15834679.1A patent/EP3234194B1/en active Active
Also Published As
Publication number | Publication date |
---|---|
EP3034621A1 (en) | 2016-06-22 |
WO2016099275A1 (en) | 2016-06-23 |
EP3234194B1 (en) | 2020-09-09 |
US20170356913A1 (en) | 2017-12-14 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Ribaut et al. | Small biomolecule immunosensing with plasmonic optical fiber grating sensor | |
EP3234194B1 (en) | New method for detecting human butyrylcholinesterase | |
CN102713578B (en) | Method for detection of an analyte in a fluid sample | |
Zhang et al. | Sensitive detection of carcinoembryonic antigen in exhaled breath condensate using surface acoustic wave immunosensor | |
US20070014692A1 (en) | Method and apparatus for measurement of binding between a protein and a nucleotide | |
WO2010080710A2 (en) | Sample collection and measurement in a single container by back scattering interferometry | |
Díaz et al. | Sol-gel cholinesterase biosensor for organophosphorus pesticide fluorimetric analysis | |
Sun et al. | Label-free detection of breast cancer biomarker using silica microfiber interferometry | |
US20120142017A1 (en) | Biosensor device and manufacturing method thereof | |
WO2004081572A1 (en) | Surface initiated thin polymeric films for chemical sensors | |
Sypabekova et al. | Ultralow limit detection of soluble HER2 biomarker in serum with a fiber-optic ball-tip resonator assisted by a tilted FBG | |
US10324034B2 (en) | Self-referencing localized plasmon resonance sensing device and system thereof | |
EP4127714A1 (en) | Rapid multiplexed serological test | |
JP2014178151A (en) | Microorganism detection method and device using microsphere resonance sensor | |
Kim et al. | Fluorescence Immunoassay of Human D‐dimer in Whole Blood | |
JP2017166825A (en) | High sensitivity detecting method and device for microsphere resonance sensors | |
US6480638B1 (en) | Single mode fiber optic evanescent wave refractometer | |
EP3612102B1 (en) | Device for detecting and/or determining the concentration of an analyte present in a tissue and a method and use of this device | |
Grime | Biochemical and clinical analysis by enthalpimetric measurements—a realistic alternative approach? | |
JP2006504950A (en) | Sensor device for determining protein aggregation | |
WO2007016665A2 (en) | Single use fluorescent assays for determination of analytes | |
CN114624196A (en) | Biosensing detection method and system for neutralizing antibody | |
WO2011123029A1 (en) | Method for determination of binding stoichiometry | |
US20160116463A1 (en) | Method for measuring the plasma concentration of an analyte directly on a whole blood sample | |
Iqbal et al. | Silicon photonic micro-ring resonators for drug screening and kinetic analysis |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE |
|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE |
|
17P | Request for examination filed |
Effective date: 20170710 |
|
AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR |
|
AX | Request for extension of the european patent |
Extension state: BA ME |
|
DAV | Request for validation of the european patent (deleted) | ||
DAX | Request for extension of the european patent (deleted) | ||
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: EXAMINATION IS IN PROGRESS |
|
17Q | First examination report despatched |
Effective date: 20180809 |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R079 Ref document number: 602015058930 Country of ref document: DE Free format text: PREVIOUS MAIN CLASS: C12Q0001680000 Ipc: G01N0033573000 |
|
GRAP | Despatch of communication of intention to grant a patent |
Free format text: ORIGINAL CODE: EPIDOSNIGR1 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: GRANT OF PATENT IS INTENDED |
|
RIC1 | Information provided on ipc code assigned before grant |
Ipc: G01N 33/573 20060101AFI20200331BHEP |
|
INTG | Intention to grant announced |
Effective date: 20200416 |
|
GRAS | Grant fee paid |
Free format text: ORIGINAL CODE: EPIDOSNIGR3 |
|
GRAA | (expected) grant |
Free format text: ORIGINAL CODE: 0009210 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE PATENT HAS BEEN GRANTED |
|
AK | Designated contracting states |
Kind code of ref document: B1 Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR |
|
REG | Reference to a national code |
Ref country code: GB Ref legal event code: FG4D |
|
REG | Reference to a national code |
Ref country code: AT Ref legal event code: REF Ref document number: 1312227 Country of ref document: AT Kind code of ref document: T Effective date: 20200915 Ref country code: CH Ref legal event code: EP |
|
REG | Reference to a national code |
Ref country code: IE Ref legal event code: FG4D |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R096 Ref document number: 602015058930 Country of ref document: DE |
|
REG | Reference to a national code |
Ref country code: LT Ref legal event code: MG4D |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: FI Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 Ref country code: SE Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 Ref country code: NO Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20201209 Ref country code: GR Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20201210 Ref country code: BG Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20201209 Ref country code: HR Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 Ref country code: LT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 |
|
REG | Reference to a national code |
Ref country code: AT Ref legal event code: MK05 Ref document number: 1312227 Country of ref document: AT Kind code of ref document: T Effective date: 20200909 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: PL Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 Ref country code: LV Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 Ref country code: RS Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: SM Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 Ref country code: PT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20210111 Ref country code: RO Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 Ref country code: EE Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 Ref country code: CZ Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: IS Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20210109 Ref country code: AL Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 Ref country code: AT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 Ref country code: ES Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R097 Ref document number: 602015058930 Country of ref document: DE |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: SK Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 |
|
PLBE | No opposition filed within time limit |
Free format text: ORIGINAL CODE: 0009261 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT |
|
REG | Reference to a national code |
Ref country code: CH Ref legal event code: PL |
|
26N | No opposition filed |
Effective date: 20210610 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: SI Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 Ref country code: DK Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 Ref country code: MC Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 |
|
REG | Reference to a national code |
Ref country code: BE Ref legal event code: MM Effective date: 20201231 |
|
REG | Reference to a national code |
Ref country code: NL Ref legal event code: FP |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: LU Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20201218 Ref country code: IE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20201218 Ref country code: IT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: CH Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20201231 Ref country code: LI Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20201231 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: TR Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 Ref country code: MT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 Ref country code: CY Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: MK Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200909 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: BE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20201231 |
|
P01 | Opt-out of the competence of the unified patent court (upc) registered |
Effective date: 20230522 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: GB Payment date: 20231220 Year of fee payment: 9 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: NL Payment date: 20231220 Year of fee payment: 9 Ref country code: FR Payment date: 20231221 Year of fee payment: 9 Ref country code: DE Payment date: 20231214 Year of fee payment: 9 |